1. Neuroscience

Visualizing endogenous opioid receptors in living neurons using ligand-directed chemistry

  1. Seksiri Arttamangkul  Is a corresponding author
  2. Andrew Plazek
  3. Emily J Platt
  4. Haihong Jin
  5. Thomas F Murray
  6. William Birdsong
  7. Kenner C Rice
  8. David Farrens
  9. John T Williams
  1. Oregon Health and Science University, United States
  2. Creighton University, United States
  3. National Institute on Drug Abuse/National Institute on Alcohol Abuse and Alcoholism, United States
https://doi.org/10.7554/eLife.49319
Share this article
Cite this article
  1. Seksiri Arttamangkul
  2. Andrew Plazek
  3. Emily J Platt
  4. Haihong Jin
  5. Thomas F Murray
  6. William Birdsong
  7. Kenner C Rice
  8. David Farrens
  9. John T Williams
(2019)
Visualizing endogenous opioid receptors in living neurons using ligand-directed chemistry
eLife 8:e49319.
https://doi.org/10.7554/eLife.49319
  2. Download BibTeX
  3. Download .RIS

Abstract

Identifying neurons that have functional opioid receptors is fundamental for the understanding of the cellular, synaptic and systems actions of opioids. Current techniques are limited to post hoc analyses of fixed tissues. Here we developed a fluorescent probe, naltrexamine-acylimidazole (NAI), to label opioid receptors based on a chemical approach termed 'traceless affinity labeling'. In this approach, a high affinity antagonist naltrexamine is used as the guide molecule for a transferring reaction of acylimidazole at the receptor. This reaction generates a fluorescent dye covalently linked to the receptor while naltrexamine is liberated and leaves the binding site. The labeling induced by this reagent allowed visualization of opioid-sensitive neurons in rat and mouse brains without loss of function of the fluorescently labeled receptors. The ability to locate endogenous receptors in living tissues will aid considerably in establishing the distribution and physiological role of opioid receptors in the CNS of wild type animals.

Data availability

All data generated or analysed during this study are includes in the manuscript and supporting files.

Article and author information

Author details

  1. Seksiri Arttamangkul

    Vollum Institute, Oregon Health and Science University, Portland, United States
    For correspondence
    arttaman@ohsu.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8815-5124

  2. Andrew Plazek

    Medicinal Chemistry Core, Oregon Health and Science University, Portland, United States
    Competing interests
    The authors declare that no competing interests exist.

  3. Emily J Platt

    Department of Biochemistry and Molecular Biology, Oregon Health and Science University, Portland, United States
    Competing interests
    The authors declare that no competing interests exist.

  4. Haihong Jin

    Medicinal Chemistry Core, Oregon Health and Science University, Portland, United States
    Competing interests
    The authors declare that no competing interests exist.

  5. Thomas F Murray

    Department of Pharmacology, School of Medicine, Creighton University, Omaha, United States
    Competing interests
    The authors declare that no competing interests exist.

  6. William Birdsong

    Vollum Institute, Oregon Health and Science University, Portland, United States
    Competing interests
    The authors declare that no competing interests exist.

  7. Kenner C Rice

    Drug Design and Synthesis Section, Intramural Research Program, National Institute on Drug Abuse/National Institute on Alcohol Abuse and Alcoholism, Bethesda, United States
    Competing interests
    The authors declare that no competing interests exist.

  8. David Farrens

    Department of Biochemistry and Molecular Biology, School of Medicine, Oregon Health and Science University, Portland, United States
    Competing interests
    The authors declare that no competing interests exist.

  9. John T Williams

    Vollum Institute, Oregon Health and Science University, Portland, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0647-6144

Funding

National Institute on Drug Abuse (DA008163)

  • John T Williams

National Institute on Drug Abuse (DA048136)

  • John T Williams

National Institute on Drug Abuse (DA042779)

  • William Birdsong

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: All animal uses were conducted in accordance with the National Institutes of Health guidelines and with approval from the Institutional Animal Care and Use Committee (IACUC) protocol #IP00000160 of the Oregon Health & Science University. Rats and mice were anesthetized with isofluorane before euthanized with minimal suffering.

Copyright

This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

Metrics

  • 4,888
    views
  • 633
    downloads
  • 43
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Seksiri Arttamangkul
  2. Andrew Plazek
  3. Emily J Platt
  4. Haihong Jin
  5. Thomas F Murray
  6. William Birdsong
  7. Kenner C Rice
  8. David Farrens
  9. John T Williams
(2019)
Visualizing endogenous opioid receptors in living neurons using ligand-directed chemistry
eLife 8:e49319.
https://doi.org/10.7554/eLife.49319

Share this article

https://doi.org/10.7554/eLife.49319

Categories and tags

Research organisms

Further reading

    1. Neuroscience

    Rhythmic circuit function is more robust to changes in synaptic than intrinsic conductances

    Zachary Fournier, Leandro M Alonso, Eve Marder
    Research Article

    Circuit function results from both intrinsic conductances of network neurons and the synaptic conductances that connect them. In models of neural circuits, different combinations of maximal conductances can give rise to similar activity. We compared the robustness of a neural circuit to changes in their intrinsic versus synaptic conductances. To address this, we performed a sensitivity analysis on a population of conductance-based models of the pyloric network from the crustacean stomatogastric ganglion (STG). The model network consists of three neurons with nine currents: a sodium current (Na), three potassium currents (Kd, KCa, KA), two calcium currents (CaS and CaT), a hyperpolarization-activated current (H), a non-voltage-gated leak current (leak), and a neuromodulatory current (MI). The model cells are connected by seven synapses of two types, glutamatergic and cholinergic. We produced one hundred models of the pyloric network that displayed similar activities with values of maximal conductances distributed over wide ranges. We evaluated the robustness of each model to changes in their maximal conductances. We found that individual models have different sensitivities to changes in their maximal conductances, both in their intrinsic and synaptic conductances. As expected, the models become less robust as the extent of the changes increases. Despite quantitative differences in their robustness, we found that in all cases, the model networks are more sensitive to the perturbation of their intrinsic conductances than their synaptic conductances.

    1. Neuroscience

    Cognition: When working memory works for our goals

    Jacob A Miller
    Insight

    When navigating environments with changing rules, human brain circuits flexibly adapt how and where we retain information to help us achieve our immediate goals.

    1. Neuroscience

    Stimulus representation in human frontal cortex supports flexible control in working memory

    Zhujun Shao, Mengya Zhang, Qing Yu
    Research Article

    When holding visual information temporarily in working memory (WM), the neural representation of the memorandum is distributed across various cortical regions, including visual and frontal cortices. However, the role of stimulus representation in visual and frontal cortices during WM has been controversial. Here, we tested the hypothesis that stimulus representation persists in the frontal cortex to facilitate flexible control demands in WM. During functional MRI, participants flexibly switched between simple WM maintenance of visual stimulus or more complex rule-based categorization of maintained stimulus on a trial-by-trial basis. Our results demonstrated enhanced stimulus representation in the frontal cortex that tracked demands for active WM control and enhanced stimulus representation in the visual cortex that tracked demands for precise WM maintenance. This differential frontal stimulus representation traded off with the newly-generated category representation with varying control demands. Simulation using multi-module recurrent neural networks replicated human neural patterns when stimulus information was preserved for network readout. Altogether, these findings help reconcile the long-standing debate in WM research, and provide empirical and computational evidence that flexible stimulus representation in the frontal cortex during WM serves as a potential neural coding scheme to accommodate the ever-changing environment.